A comprehensive characterization of single-copy T-DNA insertions in the Arabidopsis thaliana genome

被引:146
作者
Forsbach, A
Schubert, D
Lechtenberg, B
Gils, M
Schmidt, R
机构
[1] Max Planck Inst Mol Pflanzenphysiol, D-14476 Golm, Germany
[2] Icon Genet GmbH, Biozentrum Halle, D-06120 Halle An Der Saale, Germany
关键词
Agrobacterium tumefaciens; Arabidopsis thaliana; chromosome sequence map; heterochromatin; single-copy T-DNA insertion; T-DNA flanking sequence; translocation;
D O I
10.1023/A:1023929630687
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
T-DNA flanking sequences were isolated from 112 Arabidopsis thaliana single-copy T-DNA lines and sequence mapped to the chromosomes. Even though two T-DNA insertions mapped to a heterochromatic domain located in the pericentromeric region of chromosome I, expression of reporter genes was detected in these transgenic lines. TDNA insertion did not seem to be biased toward any of Arabidopsis' five chromosomes. The observed distribution of T-DNA copies in intergenic sequence versus gene sequence (i.e. 5'-upstream regions, coding sequences and 3'-downstream regions) appeared randomly. An evaluation of T-DNA insertion frequencies within gene sequence revealed that integration into 5'-upstream regions occurred more frequently than expected, whereas insertions in coding sequences (exons and introns) were found less frequently than expected based on random distribution predictions. In the majority of cases, single-copy T-DNA insertions were associated with small or large rearrangements such as deletions and/or duplications of target site sequences, deletions and/or duplications of T-DNA sequences, and gross chromosomal rearrangements such as translocations. The accuracy of integration was similarly high for both left- and right-border sequences. These results may be called upon when making detailed molecular analyses of transgenic plants or T-DNA induced mutants.
引用
收藏
页码:161 / 176
页数:16
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